豬內源性反轉錄病毒重組殼蛋白的製備與應用

Preparation and Application of the Gag Recombinant Protein of Porcine Endogenous Retrovirus

張荏婷(Jen-Ting Chang) ； 林孟雪(Meng-Shiue Lin) ； 王仕蓉(Shih-Rong Wang)

《臺灣獸醫學雜誌》 30卷1期 (2004/03) Pp. 47-55

https://doi.org/10.7009/TVJ.200403.0047

摘要

豬內源性反轉錄病毒（porcine endogenous retrovirus，PERV）經體外測試具有感染人類細胞的可能，因而造成異種移植之一大疑慮。針對此病毒之研究，我們已成功地選殖出殼蛋白基因（gag）表現質體pETST-GAG-NB3及pETST-GAG-CB7，並利用大腸桿菌之表現載體系統大量誘導出重組殼蛋白ST-GAG-NB3及ST-GAG-CB7，其大小分別為32及43 kD。我們利用此抗原進行特定豬群之篩檢，結果顯示在28頭無特定病源豬（SPF,specific pathogen-free）之血清中均無PERV抗體的反應；並且進行豬場工作員之PERV感染調查，結果顯示在34位人員中均無PERV抗體反應。此外，我們已成功的製備出殼蛋白之抗血，且經西方免疫轉漬分析證實此抗血清可與殼蛋白質結合，其大小分別為65、27及12kD。未來將可利用此重組蛋白及抗血清開發出PERV血清學檢測試劑以應用於接受異種移植病患之追蹤篩檢及PERV流行病學之研究與診斷。

關鍵字

抗血清 ； 殼蛋白 ； 豬內源性反轉錄病毒 ； 重組蛋白

並列摘要

The xenotransplantation of pig organs may be associated with a risk of transmission of microorganisms. Porcine endogenous retroviruses (PERV) are of particular concern since in vitro experiments have demonstrated that human cells are susceptible to such microorganisms. To monitor the transmission of PERV, highly sensitive and specific immunoassays must be developed for clinical surveillance. This report describes the production, preliminary characterization and application of antiserum against the recombinant PERV group specific antigens (Gag). The generated antiserum was tested using recombinant PERV Gag protein expressed in E. coli, purified PERV virus particles and human 293 cells infected with PERV. PERV translated proteins of 12.27 and 65 kilodaltons were recognized specifically by specific antiserum. Using Western blotting assay, neither 28 specific-pathogen-free pigs nor 34 farm workers could be detected antibody reaction against PERV. This work reveals that recombinant PERV Gag protein and antiserum may be effective in detecting antibodies against PERV for patients treated with xeno-transplantation, or for butchers who have extensive contact with living porcine material.

並列關鍵字

Antiserum ； Gag ； PERV ； Recombinant protein